CN102511220B - Method for measuring activity of seeds of wheat based on microscopic dynamic ion flow detecting technology - Google Patents
Method for measuring activity of seeds of wheat based on microscopic dynamic ion flow detecting technology Download PDFInfo
- Publication number
- CN102511220B CN102511220B CN 201110364011 CN201110364011A CN102511220B CN 102511220 B CN102511220 B CN 102511220B CN 201110364011 CN201110364011 CN 201110364011 CN 201110364011 A CN201110364011 A CN 201110364011A CN 102511220 B CN102511220 B CN 102511220B
- Authority
- CN
- China
- Prior art keywords
- wheat
- seeds
- wheat seed
- activity
- seed
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Landscapes
- Investigating Or Analysing Biological Materials (AREA)
- Pretreatment Of Seeds And Plants (AREA)
Abstract
The invention provides a method for measuring the activity of the seeds of wheat based on a microscopic dynamic ion flow detecting technology, which is used for carrying out dynamic K+ flow detection on the seeds of wheat at a continuous sprouting period by utilizing a microscopic dynamic ion flow detecting technology and measuring the activity of the seeds according to the net K+ flow speeds andthe directions of the seeds of the wheat. According to the method for measuring the activity of the seeds of the wheat based on the microscopic dynamic ion flow detecting technology, the non-destructive, in-vivo and quick detection of the activity of the seeds of the wheat can be realized, the time of at least several minutes and at most tens of minutes is consumed for detecting a specimen, the detecting accuracy is high, the consumed time is short, and the action of the K+ outer flows of the seeds of wheat with strong activity is weak; moreover, the action of the K+ outer flows of the seeds of wheat with weak activity is strong and is in a leaking state, the evaluating method is simple and reliable, and a non-destructive and quick novel method for detecting the activity of the seeds of wheat is provided to the seed breeding of the wheat, the storage of the seeds of the wheat and the field production of crops; and according to the novel method, a biological material after being detected still can continuously grow.
Description
Technical field
The present invention relates to the assay method of seed vitality, specifically, relate to a kind of method that flows the mensuration wheat seed vigor of detection technique based on the Micro dynamic ion.
Background technology
Seed vitality is the important indicator of seed quality, and AOSA had defined seed vitality in 1980: widely under the field condition, determine seed neatly to emerge rapidly and grow into the general name of the potential ability of seedling.ISTA classifies the standardization flow process of soya seeds accelerated aging test and pea seed leachate electrical conductivity as international seed vitality examination criteria.Current seed vitality detection method comprises: physiological measurement method, accelerated ageing method, conductimetric method, biochemical assay etc.
Micro dynamic ion stream detection technique is a kind of in a new technology that obtains lewis' acid flowing information inside and outside the test material film under the prerequisite of not destroying tested material, this technology can be measured different parts and the same position of test material continuous time, is applied in various plants research fields such as plant anti-salt research, plant pathology research, the anti-heavy metals of plant.Shabala(2000) utilize nutrient absorption during Micro dynamic ion stream detection technique has been studied wheat seeds sprouting, found K
+Stream mode outside the different parts of wheat seed all is in has confirmed K
+Outflow be since cell membrane on K
+The result of channeling is oozed out (Shabala, S. but not plant skin; Newman, I.; Wilson, S.; Clark, the ion on R.Nutrient uptake patterns over the surface of germinating wheat seeds(wheat seeds sprouting surface absorbs collection of illustrative plates), Australian Journal of Plant Physiology, 2000,27(2), 89-97), for the new visual field has been expanded in the research of wheat germination Physiology and biochemistry mechanism, at the plant anti-salt Journal of Sex Research, preventing from heavy metal is coerced this method of research field and has also been brought into play important function in addition.
Zhang Hui etc. (1996) utilize conductimetric method to measure rice paddy seed vigor (Zhang Hui, Wang Huixian, the exosmose correlation research of ion and percentage of seedgermination of rice paddy seed soak, University Of Jishou's journal, 1996,17(2): 80-83), Du Qingfu etc. (2007) have used standard sproutung method, conductimetric method, TTC sizing technique, mda content determination method mensuration corn seed vigor (Du Qingfu etc., dissimilar corn seed vigor detect the research of proper method, the corn science, 2007,15(6): 122-127).
Existing seed vitality assay method, all seed has been constituted serious damage as physiological measurement method, accelerated ageing method, conductimetric method, biochemical assay, or even destroyed kernel texture, make it lose germinating capacity fully, the loss concerning the few wheat seed of wheat seed, the quantity of preciousness of these seed vitality assay methods is quite big, and while can't realize the detection of dynamic of seed vitality.Therefore, find a kind of harmless, fast, the material after detecting can also continued growth wheat seed vigor detection method become problem demanding prompt solution.
Summary of the invention
The present invention be intended to for wheat breeding, wheat seed storage, crop field produce provide a kind of harmless, fast, biomaterial after detecting can also continued growth the wheat seed vigor detect new method.
In order to realize the object of the invention, of the present inventionly a kind ofly measure the method for wheat seed vigor based on Micro dynamic ion stream detection technique, it is to utilize Micro dynamic ion stream detection technique that the wheat seed of continuous germination period is dynamic K
+Stream detects, according to the clean K of wheat seed
+Flow velocity and direction are measured seed vitality.Wherein, grow radicle behind the described wheat seeds sprouting.
The present invention can realize harmless, live body, the fast detecting to the wheat seed vigor, and it is consuming time less to a few minutes as many as tens minutes, detection accuracy height, weak point consuming time, energetic wheat seed K to detect sample
+A little less than the outflow effect; And the weak wheat seed K of vigor
+The outflow effect is strong, presents attrition status, and evaluation method is simple and reliable.
Aforesaid method, the test ion concentration in the described Micro dynamic ion stream detection technique in the assay buffer is 0.05～0.15mM.
Particularly, preceding method may further comprise the steps: (1) pours into ion filling liquid to being full of described eletrode tip 1～1.5cm in microelectrode, the front end with electrode sucks corresponding test ion exchanger again; (2) will be inserted in the Ag/AgCl electrode wires pedestal of chlorination through the electrode after step (1) processing, and put into correcting fluid and proofread and correct; (3) get the wheat seed to be measured of continuous germination period, wheat seed radicle portion is placed on earlier in the assay buffer about balance 30min, detect 10～16min with the surface, electrode pair wheat seed radicle district to be measured after proofreading and correct again; (4) testing result is handled and analyzed.
Wherein, the described filling liquid of step (1) is: 80～140mM KCl.
The described correcting fluid of step (2) is 0.05～0.15mM KCl and 0.5～1.5mM KCl.
The described assay buffer of step (3) is: 0.05～0.15mM KCl, 0.05～0.15mMCaCl
2, 0.05～0.15mM MgCl
2, 0.3～0.6mM NaCl, 0.1～0.3mM Na
2SO
4With 0.2～0.5mM MES.
The length that the front end of step in the preceding method (1) electrode sucks the test ion exchanger is K
+: 100～200 μ m.
The Nernst equation calculating ideal value of proofreading and correct rear electrode in the preceding method is K
+: 56～60mV.
K described in the preceding method
+The flow velocity of ion stream is: sprout 32～40 hours K
+The ion internal flow velocity is-4.76～-274.99pmol.cm
-2.s
-1
The present invention can realize harmless, live body, the fast detecting to the wheat seed vigor, and it is consuming time less to a few minutes as many as tens minutes, detection accuracy height, weak point consuming time, energetic wheat seed K to detect sample
+A little less than the outflow effect; And the weak wheat seed K of vigor
+The outflow effect is strong, presents attrition status, and evaluation method is simple and reliable.
The present invention utilizes Micro dynamic ion stream detection technique to detect the K of the wheat root position of different germination periods
+Wheat seed a little less than the stream, more energetic and vigor is to K
+Hold facility is judged the vigor of wheat seed, for wheat breeding, wheat seed storage, crop field produce provide a kind of harmless, fast, biomaterial after detecting can also continued growth the wheat seed vigor detect new method.
Description of drawings
Fig. 1 the present invention is based on the experimentation figure that Micro dynamic ion stream detection technique is measured the wheat seed vigor.
Fig. 2 is K during the wheat seeds sprouting of different vigor
+Flowable state changes.
Fig. 3 is the clean K of the wheat seed of different vigor
+Flow velocity.
Figure 4 shows that when utilizing microelectrode to detect the position of the electrode distance tip of a root.
Embodiment
Following examples are used for explanation the present invention, but are not used for limiting the scope of the invention.If do not specialize the conventional means that used technological means is well known to those skilled in the art among the embodiment, the raw materials used commercial goods that is.Wherein, wheat seed is selected China spring (Triticum aestivum) for use, preserves the center available from national crops germplasm.
Embodiment
1. experimental technique
Experiment material and condition of culture: wheat seed selects for use China spring (Triticum aestivum) to be test material, liquid nutrient medium is with reference to the Hogland nutrient solution prescription, select the external form homogeneous, full wheat seed, running water cleans the back with 75% alcohol immersion 30 seconds, distilled water cleans 3-4 time then, 3% clorox soaked 10 minutes, the back is cleaned 3-4 time with distilled water, wheat seed is seeded in the culture dish of the double-deck filter paper of heelpiece, filter paper is fully wetting, and capping places 25 ± 1 ℃ incubator to sprout, surveyed primary ions stream in per 4 hours, each 10-15min.
2. experimental instrument and consumptive material:
Ion stream detects with non-damage micrometering system (BIO-001B, Younger USA Sci.﹠amp; Tech.Corp., USA), systems soft ware imFlux, it is the glass microelectrode of 2-4 μ m that microelectrode adopts tip diameter.
3. experiment reagent and solution:
(1) liquid ion exchanger comprises:
K
+: article No. Potassium ionophore I-cocktail A; Sigma-Aldrich, Louis, MO63103, USA;
(2) assay buffer composition:
K
+Ion: 0.1mM KCl, 0.1mM CaCl
2, 0.1mM MgCl
2, 0.5mM NaCl, 0.2mM Na
2SO
4, 0.3mM MES
(3) electrode filling liquid: (this is the concentration of fixing, can be influential after the change)
K
+：100mM?KCl
(4) correcting fluid:
K
+Ion: 0.1mM and 1mM KCl
4. experiment content and method:
Experimentation as shown in Figure 1.
Filling liquid to the eletrode tip that pours into the corresponding test ion about 1cm from electrode end is full of, and front end sucks the ion-exchanger (LIX) of suitable length 180-200 μ m.Electrode is inserted in the Ag/AgCl electrode wires pedestal of chlorination.Reference electrode is the low seepage electrode (WPI) of solid.Electrode is proofreaied and correct in corresponding correcting fluid, can this spy (Nerst) equation calculating ideal value be 27-30mV.
Material balance 30-50min in corresponding test fluid before the test.Then in the local vibration detection (Fig. 4) of distance wheat seed radicle district surface 30 μ m, each sample detection 10-15min.
5. data are handled
Ion stream is handled based on the Fick diffusion law, and calculates by online software MageFlux-3D Ion Flux Plotting System (http://www.xuyue.net/mageflux/).By doing statistical analysis greater than 3 resulting data of independent experiment, utilize Excel2003 to analyze mapping.
6. experimental result
Result (Fig. 2 and Fig. 3) showed before wheat seed does not send out roots K
+Be in outer stream mode always, absorb moisture after 4 hours at wheat seed, wheat seed K
+Begin outflow, the wheat seed K that vigor is weak
+Outflow is obvious, and energetic wheat seed K
+The outflow amplitude is on the low side; The wheat seed outflow amplitude a little less than the vigor reaches final big value behind the moisture to 8 hour when wheat seed absorbs, and energetic wheat seed also reaches maximum, K subsequently
+The outflow amplitude all begins to diminish, but energetic wheat seed K
+The amplitude that always outflows is all little than the wheat seed a little less than the vigor, and wheat seed energetic after 36 hours grows the tip of a root earlier, beginning active absorption K
+, and the weak wheat ion stream of vigor also is in low-level outer stream mode, its K of energetic wheat seed
+The outflow effect is little, and weak its K of wheat seed of vigor
+The outflow effect is very big, illustrated that energetic wheat seed cell membrane function is very fast with regard to what recover at the sprouting initial stage, and the weak wheat seed cell membrane function of vigor recovers slow, causes a large amount of K
+Seepage, nutrition leak has finally showed weak wheat seed vigor.
Therefore, the ion stream result of different time points can be as judging the energetic weak foundation of wheat seed as can be seen by table 1, especially at 4h, 8h and 12h, and the wheat seed K that vigor is weak
+The more energetic wheat seed that runs off is a lot of greatly, therefore can be by these several time points as the energetic weak early stage judgement of seed germination; Wheat seed energetic on the 40h time point is taken root, and K
+Stream in the beginning, the wheat seed K that vigor is weak
+Still be in outer stream mode, this time point also can be used as basis for estimation.
The wheat seed K that table 1 different time points vigor is different
+Net flow speed
Annotate: data unit is pmol/cm
2S
K
+As main nutrient ions, also play the osmotic adjustment effect, the report of relevant this respect is few, and wheat seed has been accumulated a large amount of K in maturation
+, these K
+Main nutriment as wheat seeds sprouting plays an important role in osmotic adjustment and metabolic process, and this method has found that by detecting energetic wheat seed and the wheat seed a little less than the vigor the former has very strong K
+Hold facility, and latter K
+A large amount of seepages cause nutrition leak, and basic reason is the decline of cell membrane vigor, therefore, passes through K
+Flow measuring method judge the wheat seed vigor be a kind of directly and effective method.
Though above the present invention is described in detail with a general description of the specific embodiments, on basis of the present invention, can make some modifications or improvements it, this will be apparent to those skilled in the art.Therefore, these modifications or improvements all belong to the scope of protection of present invention without departing from theon the basis of the spirit of the present invention.
Claims (4)
1. the method based on Micro dynamic ion stream detection technique mensuration wheat seed vigor is characterized in that, utilizes Micro dynamic ion stream detection technique that the wheat seed of continuous germination period is dynamic K
+Stream detects, according to the clean K of wheat seed
+Flow velocity and direction are measured seed vitality;
Grow radicle behind the described wheat seeds sprouting;
Said method comprising the steps of:
(1) pour into ion filling liquid to being full of described eletrode tip 1～1.5cm in microelectrode, the front end with electrode sucks corresponding test ion exchanger again;
(2) will be inserted in the Ag/AgCl electrode wires pedestal of chlorination through the electrode after step (1) processing, and put into correcting fluid and proofread and correct;
(3) get the wheat seed to be measured of continuous germination period, wheat seed radicle portion is placed on balance 30min in the assay buffer earlier, detect 10～16min with the surface, electrode pair wheat seed radicle district to be measured after proofreading and correct again;
(4) testing result is handled and analyzed;
Wherein, the described filling liquid of step (1) is: 80～140mM KCl;
The described correcting fluid of step (2) is 0.05～0.15mM KCl and 0.5～1.5mM KCl;
The described assay buffer of step (3) is: 0.05～0.15mM KCl, 0.05～0.15mMCaCl
2, 0.05～0.15mM MgCl
2, 0.3～0.6mM NaCl, 0.1～0.3mM Na
2SO
4With 0.2～0.5mM MES.
2. method according to claim 1 is characterized in that, the length that the front end of step (1) electrode sucks the test ion exchanger is K
+: 100～200 μ m.
3. method according to claim 1 is characterized in that, the Nernst equation calculating ideal value of proofreading and correct rear electrode is K
+: 56～60mV.
4. method according to claim 1 is characterized in that, described K
+The flow velocity of ion stream is: sprout 32～40 hours K
+The ion internal flow velocity is-4.76～-274.99pmol.cm
-2.s
-1
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201110364011 CN102511220B (en) | 2011-11-16 | 2011-11-16 | Method for measuring activity of seeds of wheat based on microscopic dynamic ion flow detecting technology |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201110364011 CN102511220B (en) | 2011-11-16 | 2011-11-16 | Method for measuring activity of seeds of wheat based on microscopic dynamic ion flow detecting technology |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102511220A CN102511220A (en) | 2012-06-27 |
| CN102511220B true CN102511220B (en) | 2013-09-04 |
Family
ID=46282529
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 201110364011 Active CN102511220B (en) | 2011-11-16 | 2011-11-16 | Method for measuring activity of seeds of wheat based on microscopic dynamic ion flow detecting technology |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102511220B (en) |
Families Citing this family (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103487482B (en) * | 2013-09-11 | 2016-02-10 | 北京农业智能装备技术研究中心 | A kind of device and using method detecting dynamic ion flow signal |
| CN103529116B (en) * | 2013-10-17 | 2015-09-30 | 北京农业信息技术研究中心 | The method and system that a kind of Heat Tolerance of Wheat Cultivars detects |
| CN104678055B (en) * | 2013-11-28 | 2016-06-29 | 旭月(北京)科技有限公司 | A kind of method differentiating gaseous safety by organism ionic molecule flow velocity |
| CN105815003B (en) * | 2016-03-23 | 2018-12-18 | 河南农业大学 | A method of seed vitality is determined based on Micro dynamic molecular flow detection technique |
| CN106076292B (en) * | 2016-06-22 | 2018-09-18 | 江西省农业科学院 | A kind of storage agent of crop seed and storage method |
| CN107094397A (en) * | 2017-04-28 | 2017-08-29 | 山东农业大学 | A kind of method of Fast nondestructive evaluation Vigor of Crop |
| CN111699782B (en) * | 2020-06-10 | 2022-02-18 | 广东省农业科学院农业生物基因研究中心 | Method for rapidly judging whether rice seeds are qualified or not by using regression equation of galactose content and germination rate |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2004034022A2 (en) * | 2002-10-08 | 2004-04-22 | Case Western Reserve University | Shape optimization to solve inverse problems and curve/model fitting problems |
-
2011
- 2011-11-16 CN CN 201110364011 patent/CN102511220B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2004034022A2 (en) * | 2002-10-08 | 2004-04-22 | Case Western Reserve University | Shape optimization to solve inverse problems and curve/model fitting problems |
Non-Patent Citations (3)
| Title |
|---|
| Nutrient uptake patterns over the surface of germinating wheat seeds;Sergey Shabala 等;《Aust.J.Plant physiol.》;20000210;第27卷(第2期);摘要第1-2行，正文左栏第3段 * |
| Sergey Shabala 等.Nutrient uptake patterns over the surface of germinating wheat seeds.《Aust.J.Plant physiol.》.2000,第27卷(第2期), |
| 张辉 等.水稻种子浸泡液外渗离子与种子发芽率的相关性研究.《吉首大学学报（自然科学版）》.1996,第17卷(第2期), * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102511220A (en) | 2012-06-27 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102511220B (en) | Method for measuring activity of seeds of wheat based on microscopic dynamic ion flow detecting technology | |
| Sun et al. | Effects of biochar, compost and straw input on root exudation of maize (Zea mays L.): From function to morphology | |
| Sayar et al. | Physiological tests as predictive appreciation for drought tolerance in durum wheat (Triticum durum Desf.) | |
| CN102393416B (en) | Method for evaluating physiological bacterial wilt of paddy rice based on non-invasive micro-detection technology | |
| Henry et al. | How does nitrogen availability alter rhizodeposition in Lolium multiflorum Lam. during vegetative growth? | |
| Jaillard et al. | Measurements of H fluxes and concentrations in the rhizosphere | |
| CN102507701B (en) | Method for screening out drought tolerant paddy rice through dynamic ion flow based on non-invasive micro-test technology (NMT) | |
| CN106034463A (en) | Method for promoting germination of oat aged seeds and growth of seedlings | |
| Mylavarapu et al. | Soil pH and Electrical Conductivity: A County Extension Soil Laboratory Manual: CIR1081/SS118, rev. 1/2020 | |
| HRP20221293T1 (en) | A method for determining microbial susceptibility to antibiotic agents | |
| CN103460852B (en) | Method for rapidly determining vitality of tobacco seeds | |
| CN102539507B (en) | Method for evaluating nitrogen nutrition of paddy by micro dynamic ion flux technique and application thereof | |
| CN110278738B (en) | Based on Ca2+Method for measuring activity of corn seeds by flow velocity | |
| Usha et al. | Evaluation of seed vigour in soybean (Glycine max) | |
| EP3845629B1 (en) | Measurement device and evaluation method | |
| Peksen et al. | Some seed traits and their relationships to seed germination and field emergence in pea (Pisum sativum L.) | |
| Farrell et al. | Net Cd2+ flux at the root surface of durum wheat (Triticum turgidum L. var. durum) cultivars in relation to cultivar differences in Cd accumulation | |
| CN103477749B (en) | A kind of Course of Corn Seed Germination phase drought resistance appraisement method based on spontaneous luminescence | |
| CN105815003B (en) | A method of seed vitality is determined based on Micro dynamic molecular flow detection technique | |
| CN109187703A (en) | A kind of copper ion selectivity micro-electrode and plant root tip copper ion measurement of rate of flow method based on non-invasive micro-test technology | |
| CN106248771A (en) | The microelectrode biosensor of a kind of original position In vivo detection Mirnas of plant and application thereof | |
| CN105445333B (en) | The method of Fast nondestructive evaluation wheat seed viability | |
| Zhu et al. | The flux rate of Ca2+ into embryo can be used to evaluate the vigour level of maize seeds | |
| Ocvirk et al. | Seed germinability after imbibition in electrical conductivity test and relations among maize seed vigour parameters | |
| CN105612851A (en) | Potassium-ion (K+)-current-based wheat salt resistance evaluation method or salt-resistant wheat screening method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |